This invention relates to a process for producing forms of magnesium ammonium phosphate (struvite) for various commercial applications to include slow-release fertilizer and possible crack repair in concrete. Fertilizer may be derived from a process for composting animal manure with a magnesium-rich compound and selected enzymes to which certain bacteria may be added to facilitate the production of a magnesium ammonium phosphate hydrate.
The formation of magnesium ammonium phosphate hexahydrate assisted by bacteria has been reported by Nabil Ben Mar et al., Chemosphere, vol. 36, No. 3, p 475-481, (1998). The removal of phosphate from supernatant liquors of anaerobically digested sludge by formation of struvite (magnesium ammonium phosphate) is reported by Battistoni et al., Wat. Res., Vol. 31, No. 11, pp. 2925-2929, 1997. The formation of struvite by Azobacter in chemically defined media has been described by Rivadeneira et al., Microbiol. 30, 55-57 (1985).
U.S. Pat. No. 6,206,945 BI, Method of Producing Artificial Guano, to Weiss et al., Mar. 27, 2001, assigned to the same assignee as this patent and incorporated herein by reference, provides a much more complicated and costly multi-step process for precipitating magnesium ammonium phosphate than the instant invention. It involves mixing manure with soft-burned dolomite at elevated temperature, recovering ammonia thus liberated in an acid, neutralizing the mixture, combining the recovered ammonium salt with the mixture, adding guano-forming bacteria, and allowing the resultant mixture to ferment.
The ""945 patent requires the following steps, each incurring expense avoided in the present invention:
a) water is added
b) carbon dioxide gas is added (and released, possibly being recaptured at additional expense)
c) high heat is generated, requiring more expensive containers
d) high alkalinity conditions require more expensive containers
e) a separate container and handling system is needed to handle the ammonia
f) acid must be supplied to mix with the ammonia
Further, work done by the present inventors indicates that magnesium ammonium phosphate need not necessarily be precipitated in large batches, but may also be suitable for xe2x80x9cgrowingxe2x80x9d in cracks in concrete. Ringelberg, David B. et al., Development of Techniques for Strengthening Deteriorated Concrete Using Bacterially-Precipitaied and Enzymatically-Precipitated Mineral Phases, 22nd Army Science Conference, Baltimore, Md., pp. 17-18, 11-13 Dec. 2000.
The combination of complementing enzymes with added bacteria in an energy efficient, simplified, cost-effective and safe process in accordance with a preferred embodiment of the present invention accelerates the production of crystalline magnesium ammonium phosphate when compared to previous methods while also increasing yield as compared to previous methods.
In general, an enhanced process produces struvite quicker and at higher yields than existing methods while employing energy efficient steps. In one embodiment, a basic form of the enhanced process yields an efficient fertilizer from animal excreta added to a mix that is then composted. The basic form of the process involves cost-effective measures of first determining the nitrogen content of the animal excreta, and then adding only that pre-specified amount of a magnesium compound to the animal excreta necessary to interact with the measured nitrogen content. Again using a cost effective means, the pH of the resultant mix is adjusted to within a pre-specified range of pH, followed by the addition of a pre-specified amount of at least one enzyme to the mix. Adding one or more enzymes increases the rate of breakdown of the urea and uric acid, thus getting ammonia production started earlier than would occur, with any existing bacteria alone. The resultant composted mix is maintained within a pre-specified temperature range, optimally requiring minimal added heat, for a pre-specified time. Pre-specified species of bacteria may then be added, providing a xe2x80x9cboosterxe2x80x9d shot of enzymes as the bacteria grow and produce their own enzymes xe2x80x9cinternally.xe2x80x9d Upon fulfilling these conditions, struvite is formed earlier in the composting process, and at higher yields, than it would be formed using processes heretofore available.
Preferred magnesium compounds include a magnesium carbonate and a magnesium hydroxide, and combinations thereof, in amounts between preferably about 1.5 and 2.0 parts, and more preferably of about 1.7 parts, of either of these magnesium compounds, or like compounds, or combinations thereof, per part of the nitrogen in the animal excreta. The enzyme, or enzymes, are added in the pre-specified amount of preferably between 0.25 and 0.75 g/l and most preferably at about 0.5 g/l (total enzyme mass) of the composted mix.
In another embodiment, the composted mix is inoculated with pre-specified species of bacteria after the enzyme (enzymes) is (are) added to the compost mix. These bacteria, through excretion of enzymes internal to them, further promote the formation of struvite as crystals outside the cell structure of the composted mix. These xe2x80x9cinternal enzymesxe2x80x9d raise the pH of the composted mix, facilitating the added bacteria to become the dominant organism in the composted mix because of the elevated pH. These pre-specified species of bacteria also contribute to maintaining high levels of the added enzyme (or enzymes) and assure that the nitrogen and phosphorous are released on an accelerated timetable and struvite formed at higher yields than previously possible.
A preferred strain of these pre-specified bacteria is the species Bacillus sphaericus. In particular, a species of Bacillus designated Bacillus sp. SB1, derived from processing bat and seabird guano, has proven to be the optimum bacteria in terms of accelerating the formation process and increasing yield of struvite. However, bacteria that have been shown to facilitate the process may be any of Bacillus sphaericus, Bacillus globisporus, and Bacillus fusiformis. 
Enzymes may be an urease, an uricase (also known as urate oxidase), intermediates such as allantoinase and allantoicase (also known as allantoine amidinohydroiase) and a phosphatase or any combination of the non-phosphatase types and phosphatase, i.e., a phosphatase and any one or more of the other enzymes. The complete pathway for the conversion of uric acid to ammonia may be described as: uric acid conversion to allantoin (due to action of uricase)xe2x80x94allantoin conversion to allantoic acid (due to action of allantoinase)xe2x80x94allantoic acid conversion to urea (due to action of allantoicase)xe2x80x94urea conversion to ammonia (due to action of urease).
A preferred urease is jack bean urease. A preferred phosphatase enzyme is a 1000 units/microliter (xcexcl) concentration of bovine alkaline phosphatase added to result in about 50 milliliter per liter (ml/l) of the composted mix after it has been mixed therein.
The pH may be adjusted within a preferred range of about 7-10 by simple aeration of the composted mix, while the mixture is maintained within a preferred temperature range of about 20-30xc2x0 C. for a preferred period of about 10-20 days and more preferably for about 14 days.
Also provided as an embodiment of the present invention is a slow-release fertilizer that efficiently provides nitrogen and phosphorous as nutrients to plants. The fertilizer is produced, at least in part, by mixing animal excreta; a pre-specified amount of a magnesium compound, and a pre-specified amount of at least one enzyme, to comprise a composted mix having a cell structure. The composted mix is then maintained within a pre-specified range of pH and a pre-specified temperature range for a pre-specified time. As a result of this process, nitrogen and phosphorous precipitate from the composted mix in a precipitate comprising at least struvite. The preferred pH range for producing this fertilizer is about 7-10. The preferred temperature range at which the composted mix is maintained is about 20-30xc2x0 C. The preferred period of composting is a range of about 10-20 days, more preferably about 14 days. This slow-release fertilizer releases nitrogen and phosphorous for use by plants over an extended period of time, with less loss to leaching than conventional fertilizers or even conventional slow-release fertilizers.
The process for producing this slow-release fertilizer may be facilitated, at least in part, by further adding bacteria to the composted mix. These bacteria, through excretion of enzymes internal to them, promote the formation of struvite as crystals outside the cell structure of the composted mix. These xe2x80x9cinternal enzymesxe2x80x9d raise the pH of the composted mix, facilitating the added bacteria in becoming the dominant organism in the composted mix. This leads to accelerated production of the fertilizer at higher yields than otherwise possible using existing methods. Further, these bacteria are added to maintain higher levels of at least one enzyme and assure that nitrogen and phosphorous are released and at least struvite is formed at an accelerated rate and higher yields than heretofore possible.
To produce this slow-release fertilizer, a preferred strain of bacteria is the species Bacillus sphaericus. In particular, a species designated Bacillus sp. SB1, derived from processing bat and seabird guano, has proven to be the optimum bacteria in terms of accelerating the formation process and increasing yield of struvite. However, bacteria that have been shown to facilitate the process may be any of Bacillus sphaericus, Bacillus globisporus, and Bacillus fusiformis. 
Enzymes used in producing this slow-release fertilizer may be either an urease, an uricase, intermediates, such as allantoinase and allantoicase (also known as allantoine amidinohydrolase), phosphatase, or a combination of the two types, i.e., a phosphatase and any one or more of the other enzymes. A preferred urease is jack bean urease. A preferred phosphatase enzyme is a 1000 units/xcexcl concentration of bovine alkaline phosphatase added to result in about 50 ml per liter of the composted mix after it has been mixed therein.
In producing this slow-release fertilizer, the pH may be adjusted within a preferred range of about 7-10 by simple aeration of the composted mix, while the mixture is maintained within a preferred temperature range of about 20-30xc2x0 C. for a preferred period of about 10-20 days and more preferably for about 14 days.
In producing this slow-release fertilizer, a preferred magnesium compound may be either a magnesium hydroxide or a magnesium carbonate, or combinations thereof, in amounts between preferably about 1.5 and 2.0 parts, and more preferably of about 1.7 parts, of either of these magnesium compounds, or like compounds, or combinations thereof, per part of the nitrogen in the animal excreta. The enzyme, or enzymes, are added in the pre-specificd amount of preferably between 0.25 and 0.75 g/l and most preferably at about 0.5 grams (total enzyme mass of all enzymes added separately) per liter of the composted mix.
In summary, composting of a mix of animal manure and a magnesium-rich compound is facilitated by the addition of selected enzymes that produce ammonia from urea and uric acid (urease or uricase) and, of course, any intermediates, and form phosphates from phosphate-rich organics, such as phosphatase. The process is accelerated by then adding to the mix selected organisms that facilitate the production of magnesium ammonium phosphate hydrate by breaking down other organic compounds and producing additional enzymes that, in turn, further accelerate release of nitrogen and phosphorous and the formation of struvite. The temperature of this mixture is maintained at about 20-30xc2x0 C. and the pH is maintained between about 7-10. Urease is added to the mixture. Next, the mixture is inoculated with bacteria, preferably of the Bacillus species, such as Bacillus sphaericus, Bacillus globisporus, and Bacillus fusiformis, and more preferably a Bacillus species designated Bacillus sp. SB1. The mixture is then allowed to incubate preferably for about 10-20 days, more preferably about 14 days to form magnesium ammonium phosphate. Optionally, phosphatase, an enzyme promoting the formation of phosphate from phosphorus-rich organic compounds, is added with an urease or uricase enzyme to further increase the yield of magnesium ammonium phosphate. Advantages of the present invention include:
increased yield of ammonium magnesium phosphate per unit of composted mass;
accelerated production of ammonium magnesium phosphate;
reduced energy cost for producing ammonium magnesium phosphate;
reduced capitalization costs for equipment to produce ammonium magnesium phosphate;
increased safety to workers in producing the ammonium magnesium phosphate;
reduced need for ancillary equipment to comply with emissions regulations;
reduced need to procure and handle caustic materials; and
reduced maintenance expenses because of lowered operating temperatures and benign components used.
Animal manure is mixed with a magnesium compound having low water solubility. Alternatively, it may be mixed with an aqueous solution of a magnesium salt. The amount of nitrogen in the animal manure is measured. Magnesium carbonate or magnesium hydroxide, preferably in the amount of about 1.5-2.0 parts and more preferably in the amount of about 1.7 parts per part of nitrogen in the manure, is added to the animal manure.
Alternatively, highly water-soluble magnesium salts, such as magnesium chloride and magnesium sulfate, may be used. However, their dosage must be carefully controlled to prevent increasing the osmotic pressure of the mixture which is detrimental to the bacteria needed for carrying out the process. In addition, magnesium sulfate may generate sulfides or mercaptans, leading to an offensive odor and toxic emissions. Advantageously, a magnesium carbonate or a magnesium hydroxide provides alkaline buffering to prevent the magnesium ammonium phosphate from dissolving after it forms.
The temperature of this mixture is maintained in the range of about 20-30xc2x0 C. Preferably, the pH is adjusted to within the range of about 7-10 by aeration. Alternatively, the pH may be adjusted by the addition of an acid. Preferably, one or more enzymes, such as urease or uricase, are added to the mixture at a preferable dosage of about 0.25-0.75 g/l, and more preferably about 0.5 g of urease (or uricase) per liter of mixture. A preferred urease is bean sprout urease and a more preferred urease is Jack bean urease Type IX at 62,100 units per gram. A suitable urease is marketed by Worthington Biochemical Corporation, Lakewood, N.J. Preferably, this solid enzyme is mixed with a 0.1 molar phosphate buffer to bring the enzyme into solution.
This mixture is then inoculated with bacteria, preferably of the species Bacillus sphaericus, Bacillus globisporus, or Bacillus fusiformis, and more preferably of the species Bacillus designated Bacillus sp. SB1 that may be isolated from guano, such as may be available from bats or seabirds. Alternatively, commercially produced guano may be used. The resultant mixture is allowed to incubate, preferably for about 10-20 days, and more preferably for about 14 days, preferably at about 20-30xc2x0 C. and more preferably at about 25 to 30xc2x0 C. to form magnesium ammonium phosphate.
In an alternate preferred embodiment of the present invention, a phosphatase enzyme is added to the batch. Typically, an alkaline phosphatase, such as bovine alkaline phosphatase (Roche Molecular Biochemicals), is used. Preferably, bovine alkaline phosphatase, at a concentration of 1000 units/microliter, is added at a dosage of 50 ml to one liter of manure. Preferably, in the alternate preferred embodiment, the phosphatase enzyme is blended with the manure after the addition of the urease or uricase and before any bacterial inoculation.